EEE 529 Microsystems

Amperometric Biosensors
Agamyrat Agambayev
520112003

Contents:
1.
2.
3.
4.
5.
6.
7.

Introduction to Biosensors
Definitions
Amperometric Biosensors
Generations of Amperometric Biosensors
Performance factors
Applications
References

MULTIDISCIPLINARY NATURE OF
BIOSENSOR:

INTRODUCTION
BIOSENSORS :
Compact analytical devices that bring together the use of a
biological, a biologically-derived or a biomimic element to recognize
the analyte.

INTRODUCTION
Biosensor = biorecognation element + transducer.
Biorecognation element
a biomolecule that recognizes the target analyte
Transducer
converts the recognition event into a measurable signal
The uniqueness of a biosensor is that the two components are
integrated into one single sensor

INTRODUCTION
Biosensor = biorecognation element + transducer.

Biosensor = biorecognation element + transducer.

Transducer. A transducer should be capable of converting the
biorecognition event into a measurable signal .
Typically, this is done by measuring the change that occur in the
bioreceptor reaction.
For example, the enzyme glucose oxidase (used as a bioreceptor
in a glucose biosensor) catalyzes the following reaction:
Glucose + O2 --------> Gluconic acid + H2O2

To measure the glucose concentration, 3 different transducers can

be used:
An oxygen sensor that measures oxygen concentration
A pH sensor that measures the acid (gluconic acid) production
A peroxide sensor that measures H2O2 concentration.
Note:
An oxygen sensor is a transducer that converts oxygen concentration into
electrical current.
A pH sensor is a transducer that converts pH change into voltage change.
A peroxidase sensor is a transducer that converts peroxidase
concentration into an electrical current.

The choice of the biological recognition element is the crucial

decision that is taken when developing a novel biosensor
design.
It is important to define criteria for, for example, a suitable
redox enzyme for a specific biosensor.
Most importantly, the enzyme needs to selectively react with
the analyte of interest.
The redox potential of the primary redox center needs to be
within a suitable potential window (usually between 0.6 and
0.9 V vs. Ag/AgCl).
The enzyme needs to be stable under the operation and
storage conditions of the biosensor and should provide a
reasonable long - term stability.

Advantages and Disadvantages of using

Enzymes in sensors
ADVANTAGES

DISADVANTAGES

Highly selective

Expensive

Catalytically active

A loss activity when

improve sensitivity
Fairly fast-acting
One of the most known
biological components

immobilized on a
transducer
Tending to lose activity
after a relatively short
period time

Considerations for biosensor development

Selection of a suitable biorecognation molecule
Selection of a suitable immobilization method
Selection of a suitable transducer
Designing of biosensor considering measurement range,
linearity, and minimization of interference
Packaging of biosensor

Biosensors are classified according to the parameter that

is measured by the physicochemical transducer of the biological event as:
Optical,
Electrochemical ---- > Amperometric
Acoustic,
Thermal.
Amperometric Biosensors: the oldest ones, which have led to the higher
number of ready to- use devices, are based on the monitoring of electrontransfer processes

Redox (reduction-oxidation) reactions include

all chemical reactions in which atoms have their oxidation
state changed.

Amperometric Biosensors produce a current

proportional to the concentration of the
substance to be detected.
The most common amperometric
biosensors use the Clark Oxygen electrode.
In the glucose Amperometric Biosensor, the
Clark Oxygen electrode is separated from
glucose by a membrane, that is permeable
to oxygen.
A biocatalyst Glucose Oxidase(GOD) is
housed between this membrane and
another membrane that separates it from
the glucose.
This membrane that separates GOD and
glucose is permeable to both Oxygen and
Glucose.

In effect, the enzyme GOD is immobilized

between two membranes, the top being
permeable only to oxygen and the bottom to
both Oxygen and Glucose.
The Glucose that enters the membrane is
Oxidised in presence of the enzyme GOD,
to produce Glucuronic acid and Hydrogen
Peroxide.
glucose + O2 > glucuronic acid + H2O2
Hence the concentration of oxygen
decreases as it moves up through the
membranes to reach the cathode. This
decrease in Oxygen concentration is
reflected as a decrease in current between
the electrodes.

Alternatively,
the decrease in the concentration of
Hydrogen Peroxide can also be used to find
the concentration of glucose, by changing
the voltage applied between the electrodes
to +0.68 V relative to the Ag/AgCl electrode
causing the reactions:
Pt anode
Ag cathode

H2O2 -- > O2 + 2H+ + 2e2AgCl + 2e- -- > 2Ag0 + 2Cl-

 1st generation: the normal product of the reaction diffuses

to the transducer and causes electrical response

 1st generation: the normal product of the reaction diffuses

to the transducer and causes electrical response

 1st generation: the normal product of the reaction diffuses

to the transducer and causes electrical response

 1st generation: the normal product of the reaction diffuses

to the transducer and causes electrical response

 2nd generation:

involves specific mediators between

reaction and transducer to generate improved response

 2nd generation:

involves specific mediators between

reaction and transducer to generate improved response

 2nd generation:

involves specific mediators between

reaction and transducer to generate improved response

 2nd generation:

involves specific mediators between

reaction and transducer to generate improved response

 2nd generation:

involves specific mediators between

reaction and transducer to generate improved response

 2nd generation:

involves specific mediators between

reaction and transducer to generate improved response

 3rd generation:

reaction itself causes the response

 3rd generation:

reaction itself causes the response

 AMPEROMETRIC BIOSENSORS

Product
detected
H2O2
choline
choline oxidase
H2O2
ethanol
alcohol oxidase
formaldehyde f. dehydrogenase NADH
glucose
glucose oxidase H2O2, O2
glutamine
glutamine oxidase H2O2
glycerol
g. dehydrogenase NADH, O2
H2O2
hypoxanthine
x. oxidase
H2O2
lactate
lactate oxidase
glucoamylase,
H2O2
oligosaccharides
glucose oxidase
Substrate

phenol

Bioreceptor

polyphenol oxidasequinone

Range,
mM
500
0 - 10
10-3
0-7 g/L
0-25
4-180
1-40
0.1-2.5

PERFORMANCE FACTORS
Selectivity
Sensitivity
Accuracy
Response time
Recovery time
Lifetime

PERFORMANCE FACTORS

References

A.G. Elie,Principles of Potentiometric and Amperometric

Biosensors,University of Virginia,(2002)
P.V. Climent, M.L.M.Serralheiro, and M.J.F. Rebelo, Pure and Applied
Chemistry 73, pp.1993-1999, 2001
http://www.lsbu.ac.uk.html
http://www.gatewaycoalition.org/files/Hidden/sensr/tocsenf.htm
http://techramble.wordpress.com/2009/08/03/amperometric-biosensors/
http://www.lbb.ethz.ch/Education/Biosensors/2011FSlesson4TZ

Thank You